Inactivation of dispatched 1 by the chameleon mutation disrupts Hedgehog signalling in the zebrafish embryo
Inactivation of dispatched 1 by the chameleon mutation disrupts Hedgehog signalling in the zebrafish embryo
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DOI：
10.1016/j.ydbio.2004.01.022
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发表时间：
2004-05-13
影响因子：
2.7
通讯作者：
Ingham, PW
中科院分区：
文献类型：
--
作者：
Nakano, Y;Kim, HR;Ingham, PW
对斑马鱼EST和全基因组鸟枪序列数据库中编码甾醇敏感结构域（SSD）蛋白基序的序列进行检索，发现两组DNA序列与果蝇分泌的Hedgehog蛋白释放所需的派遣基因具有显著的同源性。使用吗啉代反义寡核苷酸，我们发现，这些基因之一，指定Disp 1的抑制，结果在一个表型类似的“你型”突变体，以前牵连在信号转导的刺猬蛋白在斑马鱼胚胎。注射disp1 mRNA到胚胎纯合的一个这样的突变，变色龙（CON）的结果在救援的突变表型。辐射杂交定位定位disp1到同一地区的LG20的映射通过减数分裂重组分析的构象。来自纯合子con突变胚胎的disp1 cDNA的序列分析显示，这两个突变等位基因与disp1编码序列中的提前终止密码子相关。通过分析特定细胞类型的标记物在神经管，胰腺和肌节的CON突变体和Disp1 morphant胚胎的表达，我们得出结论，Disp1活性是必不可少的脂质修饰的Hh蛋白质的分泌中线结构。(C)2004爱思唯尔公司All rights reserved.
Searches of zebrafish EST and whole genome shotgun sequence databases for sequences encoding the sterol-sensing domain (SSD) protein motif identified two sets of DNA sequences with significant homology to the Drosophila dispatched gene required for release of secreted Hedgehog protein. Using morpholino antisense oligonucleotides, we found that inhibition of one of these genes, designated Disp1, results in a phenotype similar to that of the "you-type" mutants, previously implicated in signalling by Hedgehog proteins in the zebrafish embryo. Injection of disp1 mRNA into embryos homozygous for one such mutation, chameleon (con) results in rescue of the mutant phenotype. Radiation hybrid mapping localised disp1 to the same region of LG20 to which the con mutation was mapped by meiotic recombination analysis. Sequence analysis of disp1 cDNA derived from homozygous con mutant embryos revealed that both mutant alleles are associated with premature termination codons in the disp1 coding sequence. By analysing the expression of markers of specific cell types in the neural tube, pancreas and myotome of con mutant and Disp1 morphant embryos, we conclude that Disp1 activity is essential for the secretion of lipid-modified Hh proteins from midline structures. (C) 2004 Elsevier Inc. All rights reserved.